Field of the Invention
Embodiments of the present disclosure relate in general to methods for determining the presence of a polymorphism at a target nucleotide position. Embodiments of the present disclosure also relate in general to methods of introducing known sequences flanking a target nucleotide position into amplicons of a target nucleic acid sequence. A probe complementary to the known sequences flanking the target nucleotide position can then be used to identify the presence of a polymorphism at the target nucleotide position.
Description of Related Art
Methods exist to identify polymorphisms in target nucleic acid sequences. See U.S. Pat. No. 7,368,242; U.S. Pat. No. 6,830,887; US 20030054339; U.S. Pat. No. 8,318,428; U.S. Pat. No. 7,115,364; U.S. Pat. No. 6,410,231; US 20120088246; US 20100009355; US 20130045881; WO1999016910; Billard et al., Appl. Environ. Microbiol. (2012); 78(4):1063-8; Hymas et al., J. Virol. Methods (2007); 142(1-2): 10-14; Rowley et al., AIDS Res. Hum. retroviruses (2010); 26(3):293-300; Rowley et al., J. Virol. methods (2008); 149(1): 69-75; Stevenson et al., J. Clin. Microbiol. (2005); 43(5):2391-98; and Whiley et al., J. Clin. Virol. (2006); 35(1): 81-83.
However, in certain replicating organisms, one or more secondary polymorphisms may be present near a primary polymorphism in a target nucleic acid sequence which may prevent or inhibit probe binding to the target probe binding site, thereby resulting in a false negative.